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Fig. 1. Characterization of FO-1 cells expressing the FcRn {alpha}-chain alone or in combination with ß2m. (A) Western blot analysis. Lysates of FO-1 cells (lane 1), FO-1 ß2m cells (lane 2), FO-1 cells expressing Myc-hFcRn (lane 3), and FO-1 ß2m cells expressing Myc-hFcRn (lane 4) were analyzed by SDS-PAGE and immunoblotting using polyclonal anti-FcRn peptide antibodies ({alpha}-FcRn; upper panel), monoclonal anti-Myc antibodies ({alpha}-Myc; middle panel), and polyclonal anti-ß2m antibodies ({alpha}-ß2m; lower panel). For reasons unknown, the anti-FcRn serum detected the {alpha}-chain less well in lysates from FO-1 ß2m than in FO-1 cells. (B) Co-localization of the FcRn {alpha}-chain and ß2m. FO-1 cells (a,c) and FO-1 ß2m cells expressing Myc-hFcRn (c,d,e) were fixed, permeabilized and stained with monoclonal anti-Myc ({alpha}-Myc; a,b) and polyclonal anti-ß2m antibodies ({alpha}-ß2-m; c,d). Panel e shows the merged staining for Myc-hFcRn (green) and ß2m (red). (C) Co-immunoprecipitation of the FcRn {alpha}-chain and ß2m. FO-1 cells (lanes 1,5), FO-1 cells expressing Myc-hFcRn (lanes 2,6), FO-1 ß2m cells (lanes 3,7) and FO-1 ß2m cells expressing Myc-hFcRn (lanes 4,8) were lyzed and immunoprecipitated with monoclonal anti-Myc antibodies ({alpha}-Myc IP; lanes 1-4) or polyclonal anti-ß2m antibodies ({alpha}-ß2m IP; lanes 5-8). Precipitated proteins were immunoblotted with polyclonal anti-Myc antibodies ({alpha}-Myc; top panel) and polyclonal anti-ß2m antibodies ({alpha}-ß2m; bottom panel). The data in Fig. 1 is representative of at least three independent experiments carried out using two different cell clones.